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Modifying current thin-film microextraction(TFME)solutions for analyzing prohibited substances:Evaluating new coatings using liquid chromatography

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Journal of Pharmaceutical Analysis 2022年第3期12卷 470-480页

作者： Łukasz Sobczak Dominika Kołodziej Krzysztof Gorynski Bioanalysis Scientific Group Faculty of PharmacyCollegium Medicum in Bydgoszcz at Nicolaus Copernicus University in Torun85-089BydgoszczPoland

For identifying and quantifying prohibited substances,solid-phase microextraction(SPME)continues to arouse interest as a sample preparation method.However,the practical implementation of this method in routine laboratory testing is currently hindered by the limited number of coatings compatible with the ubiquitous high-performance liquid chromatography(HPLC)systems.Only octadecyl(C18)and polydimethylsiloxane/divinylbenzene ligands are currently marketed for this purpose.To address this situation,the present study evaluated 12 HPLC-compatible coatings,including several chemistries not currently used in this application.The stationary phases of SPME devices in the geometry of thin filmcoated blades were prepared by applying silica particles bonded with various functional ligands(C18,octyl,phenyl-hexyl,3-cyanopropyl,benzenesulfonic acid,and selected combinations of these),as well as unbonded silica,to a metal support.Most of these chemistries have not been previously used as microextraction coatings.The 48 most commonly misused substances were selected to assess the extraction efficacy of each coating,and eight desorption solvent compositions were used to optimize the desorption conditions.All samples were analyzed using an HPLC system coupled with triple quadrupole tandem mass spectrometry.This evaluation enables selection of the best-performing coatings for quantifying prohibited substances and investigates the relationship between extraction efficacy and the physicochemical characteristics of the analytes.Ultimately,using the most suitable coatings is essential for trace-level analysis of chemically diverse prohibited substances.

关键词： Sample preparation Solid-phase microextraction Thin-film microextraction Prohibited substances Drugs of abuse High-performance liquid chromatography

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Risk of venous thromboembolic disease in postmenopausal women taking oral or transdermal hormone replacement therapy

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2011年第1期12卷 12-17页

作者： Barbara RUSZKOWSKA GraZyna GADOMSKA Liliana BIELIS Marzena GRUSZKA Barbara GóRALCZYK Danuta RO GraZyna ODROWAZ-SYPNIEWSKA Department of Pathophysiology Collegium Medicum in BydgoszczNicolaus Copernicus University in Toruń Department of Laboratory Medicine Collegium Medicum in BydgoszczNicolaus Copernicus University in Toruń Department of Laboratory Medicine Collegium Medicum in BydgoszczNicolaus Copernicus University in Toruń

Objective: The influence of hormone replacement therapy (HRT) on hemostasis processes depends on the type of hormone,the combination of doses,the time of taking HRT,and the route of administration (oral,transdermal,implanted).The aim of the current study was to assess some parameters of coagulation,especially tissue factor pathway inhibitor (TFPI) and tissue factor (TF) in postmenopausal women using oral or transdermal HRT.Methods: The study was conducted on 76 healthy women,including 46 women aged 44–58 years who were taking oral (26) or transdermal (20) HRT,and 30 women aged 44–54 years who did not take HRT as the control group.Plasma concentrations of TF,TFPI,thrombin-antithrombin complex (TAT),and D-dimer were performed by enzyme-linked immunosorbent assay (ELISA).Moreover,the concentration of fibrinogen and activity of protein C were measured by chromogenic and chronometric methods.Results: We observed a significantly higher concentration of TF and a significantly lower concentration of TFPI in women taking oral and transdermal HRT in comparison with the control group.We also found a significantly lower concentration of fibrinogen in women taking oral HRT vs.the control group.Moreover,no statistically significant changes in concentrations of TAT and D-dimer,or activity of protein C were noted.Conclusions: In this study,the occurrence of an increased TF concentration simultaneously with a decreased concentration of TFPI in women taking HRT indicates hypercoagulability.No significant modification of TAT or D-dimer occurred,and thus there may not be increased risk of thrombosis.

关键词： Menopause Coagulation Hormone replacement therapy

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Assessment of ghrelin and leptin receptor levels in postmenopausal women who received oral or transdermal menopausal hormonal therapy

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2012年第1期13卷 35-42页

作者： Barbara RUSZKOWSKA Alina SOKUP Arleta KULWAS Maciej W.SOCHA Krzysztof GóRALCZYK Barbara GóRALCZYK Danuta ROS Department of Pathophysiology Collegium Medicum in BydgoszczNicolaus Copernicus University in Toruń Department of Gastroenterology Angiology and Internal DiseasesCollegium Medicum in BydgoszczNicolaus Copernicus University in Toruń Department of Obstetrics Gynecology and Oncological GynecologyCollegium Medicum in BydgoszczNicolaus Copernicus University in Toruń

Objective:In postmenopausal women,an increased leptin concentration and reduced levels of ghrelin and adiponectin were observed.The aim of this study was to evaluate the concentrations of the active form of ghrelin,total ghrelin,leptin receptor,lipoprotein(a)(Lp(a)),and plasminogen activator inhibitor type 1(PAI-1) in postmenopausal women who received oral or transdermal menopausal hormonal therapy(MHT).Methods:The study involved 76 healthy women:46 women aged from 44 to 58 years who received oral(26) or transdermal(20) MHT;the control group consisted of 30 women aged from 44 to 54 years who did not receive MHT.The plasma concentrations of total ghrelin,the active form of ghrelin,Lp(a),and PAI-1:Ag were measured by enzyme-linked immunosorbent assay(ELISA).The concentration of the leptin receptor was measured by enzyme immunometric assay(EIA).Results:We observed a significantly higher concentration of total ghrelin and the active form of ghrelin in women who received transdermal MHT in comparison with those who took oral MHT.We also found a significantly lower concentration of total ghrelin in women who received oral MHT compared with the control group.A higher concentration of PAI-1:Ag was found in the group of women who took transdermal MHT in comparison with those who took oral MHT and with the control group.The differences were statistically significant.Additionally,we found a significant negative correlation between the concentrations of total ghrelin and PAI-1:Ag and a positive correlation between the concentrations of total ghrelin and leptin receptor in women who received transdermal MHT.Conclusions:The study showed that women who used transdermal MHT had higher levels of total ghrelin than women who took oral MHT.This indicates a beneficial effect of the transdermal route of MHT.However,transdermal therapy was associated with adverse effects with regard to the observed higher levels of PAI-1:Ag,which in turn,can lead to a reduction in fibrinolytic activity.

关键词： ghrelin 瘦素受体激素治疗绝经后妇女纤溶酶原激活物抑制物高水口服

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Solid-phase microextraction of endogenous metabolites from intact tissue validated using a Biocrates standard reference method kit

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Journal of Pharmaceutical Analysis 2023年第1期13卷 55-62页

作者： Runshan Will Jiang Karol Jaroch Janusz Pawliszyn Department of Chemistry University of WaterlooWaterlooN2L 3G1Canada Department of Pharmacodynamics and Molecular Pharmacology Faculty of PharmacyCollegium Medicum in BydgoszczNicolaus Copernicus University in TorunBydgoszcz85-089Poland

Improved analytical methods for the metabolomic profiling of tissue samples are constantly needed.Currently,conventional sample preparation methods often involve tissue biopsy and/or homogenization,which disrupts the endogenous metabolome.In this study,solid-phase microextraction(SPME)fibers were used to monitor changes in endogenous compounds in homogenized and intact ovine lung tissue.Following SPME,a Biocrates AbsoluteIDQ assay was applied to make a downstream targeted metabolomics analysis and confirm the advantages of in vivo SPME metabolomics.The AbsoluteIDQ kit enabled the targeted analysis of over 100 metabolites via solid-liquid extraction and SPME.Statistical analysis revealed significant differences between conventional liquid extractions from homogenized tissue and SPME results for both homogenized and intact tissue samples.In addition,principal component analysis revealed separated clustering among all the three sample groups,indicating changes in the metabolome due to tissue homogenization and the chosen sample preparation method.Furthermore,clear differences in free metabolites were observed when extractions were performed on the intact and homogenized tissue using identical SPME procedures.Specifically,a direct comparison showed that 47 statistically distinct metabolites were detected between the homogenized and intact lung tissue samples(P<0.05)using mixed-mode SPME fibers.These changes were probably due to the disruptive homogenization of the tissue.This study's findings highlight both the importance of sample preparation in tissue-based metabolomics studies and SPME's unique ability to perform minimally invasive extractions without tissue biopsy or homogenization while providing broad metabolite coverage.

关键词： Solid-phase microextraction Solvent extraction Metabolomics Sample preparation In vivo sampling

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Decreased serum prohepcidin concentration in patients with polycythemia vera

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2009年第11期10卷 791-795页

作者： Justyna KWAPISZ Ewa ■EKANOWSKA Joanna JASINIEWSKA Department of Pathophysiology Nicolaus Copernicus University in ToruńCollegium Medicum in Bydgoszcz

客观的铁缺乏是在有红血球增多症 vera (PV ) 的病人的普通复杂并发症。Hepcidin 是铁动态平衡的一个主要管理者。我们的学习的目的是在 PV 病人的浆液估计 prohepcidin，一位 hepcidin 先锋，和另外的铁地位参数。学习在 60 个病人(F/M... 详细信息

客观的铁缺乏是在有红血球增多症 vera (PV ) 的病人的普通复杂并发症。Hepcidin 是铁动态平衡的一个主要管理者。我们的学习的目的是在 PV 病人的浆液估计 prohepcidin，一位 hepcidin 先锋，和另外的铁地位参数。学习在 60 个病人(F/M 26/34 ) 被执行的方法变老 38 ∼84 (66 ± 1 0 ) 年。控制组由 20 个健康志愿者，匹配的年龄和性别组成了。下列参数在血浆液样品被决定：prohepcidin 集中，熨满意的、不饱和的铁有约束力的能力(UIBC ) ，全部的铁绑定能力(TIBC ) ， transferrin 浸透(TfS ) ，和含铁锡和可溶的 transferrin 受体(sTfR ) 的集中。所有 PV 病人显著地显示出的结果 prohepcidin 的底层，与控制组相比的 sTfR 和 TIBC 高级。从学习组的 40% 病人在正常范围下面并且显著地显示出含铁锡的集中浆液铁和 TfS 的底层，并且与学习组的其余部分比较的 sTfR， UIBC 和 TIBC 显著地高级。在这组病人， prohepcidin 集中是比在另外的病人的那些显著地低的。结果显示那 PV 的结论病人们受不了铁新陈代谢混乱。在 PV 病人的 prohepcidin 的减少的浆液水平可以是铁缺乏的结果。

关键词：血清浓度增多症红细胞患者转铁蛋白受体铁缺乏症抗菌多肽结合力

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Time-course monitoring of in vitro biotransformation reaction via solid-phase microextraction-ambient mass spectrometry approaches

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Journal of Pharmaceutical Analysis 2022年第1期12卷 186-191页

作者： Karol Jaroch Janusz Pawliszyn Department of Chemistry University of WaterlooWaterlooN2L 3G1Canada Department of Pharmacodynamics and Molecular Pharmacology Faculty of PharmacyCollegium Medicum in BydgoszczNicolaus Copernicus University in Toruń Poland85090BydgoszczPoland

The solid-phase microextraction technique quantifies analytes without considerably affecting the sample composition.Herein,a proof-of-concept study was conducted to demonstrate the use of coated probe electrospray ionization(coated-PESI)and coated blade spray(CBS)as ambient mass spectrometry approaches for monitoring drug biotransformation.The ability of these methods was investigated for monitoring the dephosphorylation of a prodrug,combretastatin A4 phosphate(CA4P),into its active form,combretastatin A4(CA4),in a cell culture medium supplemented with fetal bovine serum.The CBS spot analysis was modified to achieve the same extraction efficiency as protein precipitation and obtained results in 7 min.Because coated-PESI performs extraction without consuming any samples,it is the preferred technique in the case of a limited sample volume.Although coated-PESI only extracts small quantities of analytes,it uses the desorption solvent volume of 5-10 pL,resulting in high sensitivity,thus allowing the detection of compounds after only 1 min of extraction.The biotransformation of CA4P into CA4 via phosphatases occurs within the simple matrix,and the proposed sample preparation techniques are suitable for monitoring the biotransformation.

关键词： Solid-phase microextraction Coated probe electrospray ionization Coated blade spray Combretastatin Biotransformation Prodrug activation

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How rotational speed of planetary ball mill and polymer load influence the performance and water vapor sorption in solid dispersions composed of tadalafil and soluplus

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Particuology 2023年第2期73卷 37-46页

作者： Karol Kubat Anna Krupa Witold Brniak Agnieszka Węgrzyn Dorota Majda Agata Bogdał Hubert Harańczyk Department of Medical Physics Faculty of PhysicsInstitute of PhysicsJagiellonian UniversityCracowPoland Department of Pharmaceutical Technology and Biopharmaceutics Faculty of PharmacyCollegium MedicumJagiellonian UniversityCracowPoland Faculty of Chemistry Jagiellonian UniversityCracowPoland

The presence of residual water may deteriorate the performance of amorphous solid dispersions prepared by ball milling,affecting molecular mobility,crystallinity,particle size and finally,the drug dissolution rate.As the stability of these metastable systems depend on both formulation and process variables,the aim of this study was to assess for the first time,the impact that the polymer load and the rotational speed applied upon high energy ball milling could have on the performance of binary co-milled solid dispersions composed of tadalafil(a hydrophobic crystalline drug)and Soluplus(an amphiphilic,hygroscopic amorphous polymer).Each of these variables was tested at three levels.Scanning electron microscopy,laser diffraction and X-ray powder diffraction were used to analyze morphology,particle size distribution and crystallinity of ball milled formulations respectively.Dissolution studies were also carried out.Advanced tools of applied physics,namely solid state 1H NMR and relaxometry were used to assess the structure and water mobility upon gaseous phase hydration on storage.It was shown that both tested variables determined the particle size of the formulation.When the rotational speed of 400 rpm was used,all solid dispersion were XRD-amorphous,but to ensure the immediate release of tadalafil its micellar solubilization in Soluplus was necessary.While the formulation was exposed to water vapor,the hydration level increased with an increasing polymer load as well.Hence,the rotational speed governed the space available for the adsorption of water molecules and their organization in a monolayer or multilayers.Such behavior may have impact on the kinetics of the amorphous drug recrystallization,and finally deteriorate its dissolution.

关键词： Tadalafil Soluplus Solid dispersion Relaxation time Milling NMR

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Pressurized intraperitoneal aerosol chemotheprapy after misdiagnosed gastric cancer: case report and review of the literature

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World Journal of Gastroenterology 2018年第19期24卷 2130-2136页

作者： Maciej Nowacki Dariusz Grzanka Wojciech Zegarski Department of Surgical Oncology Ludwik Rydygier's Collegium Medicum in Bydgoszcz Nicolaus Copernicus University in Torun Oncology Centre-Prof. Franciszek ?ukaszczyk Memorial Hospital in Bydgoszcz Department of Clinical Pathomorphology Ludwik Rydygier's Collegium Medicum in Bydgoszcz Nicolaus Copernicus University in Torun

We report the first application of pressurized intraperitoneal aerosol chemotherapy(PIPAC) as a rescue therapy before palliative D2 gastrectomy combined with liver metastasectomy performed in a 49-yearold woman with peritoneal carcinomatosis who was primarily diagnosed with and underwent surgery for a Krukenberg tumor. The PIPAC procedure was performed with the use of cisplatin at 7.5 mg/m2 and doxorubicin at 1.5 mg/m2 for 30 min at 37 ℃. Eight weeks after the PIPAC procedure, the patient underwent open classic D2 gastrectomy with the creation of a Roux-en-Y anastomosis(RNY) combined with liver metastasectomy. The patient underwent the classic protocol for chemotherapy combined with Xeloda. The patient felt better and returned to her daily activities. Multicenter data should be gathered to confirm the usefulness of PIPAC as a rescue or neoadjuvant supportive therapy in a very select group of patients who have been recently qualified to undergo classic chemotherapy or standard oncologic surgical procedures.

关键词： Peritoneal carcinomatosis Pressurized intraperitoneal aerosol chemotherapy Neoadjuvant therapy Gastric cancer Krukenberg tumor

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Safety of intrathecal injection of Wharton's jellyderived mesenchymal stem cells in amyotrophic lateral sclerosis therapy

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Neural Regeneration Research 2019年第2期14卷 313-318页

作者： Monika Barczewska Mariusz Grudniak Stanis?aw Maksymowicz Tomasz Siwek Tomasz O?dak Katarzyna Jezierska-Wo?niak Dominika G?adysz Wojciech Maksymowicz Department of Neurology and Neurosurgery School of Medicine Collegium Medicum-University of Warmia and Mazury in Olsztyn University Clinical Hospital in Olsztyn Instytut Terapii Komórkowych w Olsztynie(Cell Therapies Institute FamiCord Group) Polski Bank Komórek Macierzystych (PBKM FamiCord Group) Department of Psychology Clinical Logopedics and Social Science in Medicine Faculty of Health Sciences Collegium Medicum-University of Warmia and Mazury in Olsztyn Department of Neurology and Neurosurgery Laboratory of Regenerative Medicine School of Medicine Collegium Medicum-University of Warmia and Mazury in Olsztyn

Animal experiments have confirmed that mesenchymal stem cells can inhibit motor neuron apoptosis and inflammatory factor expression and increase neurotrophic factor expression. Therefore, mesenchymal stem cells have been shown to exhibit prospects in the treatment of amyotrophic lateral sclerosis. However, the safety of their clinical application needs to be validated. To investigate the safety of intrathecal injection of Wharton's jelly-derived mesenchymal stem cells in amyotrophic lateral sclerosis therapy, 43 patients(16 females and 27 males, mean age of 57.3 years) received an average dose of 0.42 × 106 cells/kg through intrathecal administration at the cervical, thoracic or lumbar region depending on the clinical symptoms. There was a 2 month interval between two injections. The adverse events occurring during a 6-month treatment period were evaluated. No adverse events occurred. Headache occurred in one case only after first injection of stem cells. This suggests that intrathecal injection of Wharton's Jelly-derived mesenchymal stem cells is well tolerated in patients with amyotrophic lateral sclerosis. This study was approved by the Bioethical Committee of School of Medicine, University of Warmia and Mazury in Olsztyn, Poland(approval No. 36/2014 and approval No. 8/2016). This study was registered with the ClinicalTrials.gov(identifier: NCT02881476)on August 29, 2016.

关键词： amyotrophic lateral sclerosis stem cells therapy intrathecal injections Wharton's jelly-derived mesenchymal stem cells adverse events safety cerebrospinal fluid neural regeneration

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One extraction tool for in vitro-in vivo extrapolation?SPME-based metabolomics of in vitro 2D,3D,and in vivo mouse melanoma models

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Journal of Pharmaceutical Analysis 2021年第5期11卷 667-674页

作者： Karol Jaroch Paulina Taczynska Marta Czechowska Joanna Bogusiewicz KamilŁuczykowski Katarzyna Burlikowska Barbara Bojko Department of Pharmacodynamics and Molecular Pharmacology Faculty of PharmacyCollegium Medicum in BydgoszczNicolaus Copernicus University in Torun Poland85-089BydgoszczPoland

Solid phase microextraction(SPME)in combination with high-resolution mass spectrometry was employed for the determination of metabolomic profile of mouse melanoma growth within in vitro 2D,in vitro 3D,and in vivo models.Such multi-model approach had never been investigated before.Due to the low-invasiveness of SPME,it was possible to perform time-course analysis,which allowed building time profile of biochemical reactions in the studied material.Such approach does not require the multiplication of samples as subsequent analyses are performed from the very same cell culture or from the same individual.SPME already reduces the number of animals required for experiment;therefore,it is with good concordance with the 3Rs rule(replacement,reduction,and refinement).Among tested models,the largest number of compounds was found within the in vitro 2D cell culture model,while in vivo and in vitro 3D models had the lowest number of detected compounds.These results may be connected with a higher metabolic rate,as well as lower integrity of the in vitro 2D model compared to the in vitro 3D model resulting in a lower number of compounds released into medium in the latter model.In terms of in vitro-in vivo extrapolation,the in vitro 2D model performed more similar to in vivo model compared to in vitro 3D model;however,it might have been due to the fact that only compounds secreted to medium were investigated.Thus,in further experiments to obtain full metabolome information,the intraspheroidal assessment or spheroid dissociation would be necessary.

关键词： Solid-phase microextraction In vitro-in vivo extrapolation Metabolomics Melanoma B16F0 C57BL6 mouse model

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